黄河裸裂尻鱼ob基因克隆及环境温度对基因表达 - 中国实验动物学报.doc

黄河裸裂尻鱼基因克隆赵兰英1，晁 燕1，李长忠2，2，申志新3，王国杰3，3，李 超1，拜彬强1，祁得林1(青海省高原水生生物及生态环境重点实验室 【摘要】 目的 克隆黄河裸裂尻鱼肥胖基因（ob），分析编码蛋白leptin的序列特征，检测ob mRNA的组织特异性表达和不同环境温度下ob mRNA的相对表达水平，为探究黄河裸裂尻鱼在高原季节性寒冷环境下的生态适应机制科学数据。方法 利用RT-PCT、5′-RACE和3-RACE法获得ob基因全长cDNA序列。通过已知cDNA序列设计半定量RT-PCR和Real-time PCR引物，进行表达研究。结果 黄河裸裂尻鱼ob基因序列全长为1bp，其中开放阅读框（ORF）长5bp，编码1个氨基酸。黄河裸裂尻鱼ob mRNA在心脏肾脏、脂肪、肠、精巢肝胰脏鳃、脑心脏中表达最强，肌肉组织中表达。ob mRNA的表达。结论 黄河裸裂尻鱼ob基因特殊的组织表达特征可能与其特定组织的能量代谢及其特殊的生物学功能有关。Leptin在黄河裸裂尻鱼能量代谢调节和季节性环境适应方面发挥着重要作用。 【关键词黄河裸裂尻鱼肥胖基因克隆表达Molecular cloning of ob gene in Schizopygopsis pylzovi and its mRNA expression responding to seasonal cold environment of the Qinghai-Tibetan Plateau ZHAO Lan-ying1, CHAO Yan1, LI Chang-zhong2, XIE Bao-sheng2, SHEN Zhi-xin3, WANG Guo-jie3, WANG Zhen-gang3, LI Chao1, BAI Bin-qiang1, QI De-lin1 (1. Animal Science Department of Agriculture and Animal Husbandry College, Qinghai University, Xining 810016, China; 2. Department?of?Biological?Sciences, Qinghai?University, Xining?810016, China; 3. Key Lab for Plateau Hydrobiology and Ecological Environment of Qinghai Province, Fishery Environmental Monitoring Station of Qinghai Province, Xining 810012, China) 【Abstract】Objective In order to provide scientific data for the adaptive mechanism of Schizopygopsis pylzovi to seasonal cold environment of the Qinghai-Tibetan Plateau, the obese gene (ob) of Schizopygopsis pylzovi was cloned and its encoding protein leptin was analyzed. Then the expression pattern of the ob mRNA in tissues and the relative expression level of the ob mRNA in hepatopancreas and white muscle of Schizopygopsis pylzovi from habitats with different temperatures. Methods The full-length cDNA of ob gene was cloned by RT-PCT, 5-RACE and 3-RACE methods. The expression pattern and relative expression level were determined with RT-PCR and Real-time PCR. Results The full-length cDNA of ob gene was 1337 bp, including a 513 bp complete ORF encoding a 170 amino acid peptide. The ob gene was expressed in the nine examined tissues, includin